Marine Biological Valuation (BV) has increased in importance in recent years, due to the need to establish accurate maps of biodiversity value. However, there have been few exercises undertaken in Southern Europe, in putting a value on marine biodiversity whilst at the same time looking at several biological components. This paper presents the complete Biological Valuation Map (BVM) of the Basque continental shelf and estuaries, using the methodology developed for the Belgian Continental Shelf. It includes all available biological data (zooplankton, macroalgae, macrobenthos, demersal fish, seabirds and cetaceans), from 2003 to 2010. BVMs aim to compile all available biological and ecological information for a selected study area, allocating an integrated intrinsic biological value to the subzones within the study area. Here, the results highlight specific areas (such as Jaizkibel or Cap Breton Canyon), as having high or very high integrated BV, using all of the components. Furthermore, some biodiversity ‘hotspots’ have been identified, according to a specific ecosystem component (e.g. mid-parts of the Oka estuary, for macroalgae, and the Cap Breton Canyon, for cetaceans). Comparison with the results obtained from other European countries, and with previously high-importance delimited zones within the study area, showed similar spatial trends and patterns. Therefore, the objectives of this contribution are: (i) to analyse and establish a spatial ecological value map of the continental shelf of the Basque Country (southern Bay of Biscay), using present BV methods; (ii) to compare the results obtained to other European countries, and (iii) to explore the application of these results to the Marine Spatial Planning (MSP) and the European Marine Strategy Framework Directive (MSFD) requirements. This map can serve as a baseline for future MSP and can also be used for the determination of the environmental status, within the MSFD, for the qualitative descriptor 1 (biodiversity).